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Introduction: Electrolyte imbalance refers to altered electrolyte levels that predominantly affect diabetic patients due to hyperglycemia which rise plasma osmolality and impaired renal function, contributing to a change in electrolyte level. Therefore, this study aimed to assess the prevalence of electrolyte imbalance and its associated factors among diabetic patients and healthy control groups attending the University of Gondar Comprehensive Specialized Hospital. Patients and Methods: A comparative cross-sectional study was conducted on 130 diabetic patients and 130 diabetes-free controls. Sociodemographic, behavioral and clinical data were collected using a structured questionnaire. After measuring anthropometric parameters, 5mL of the blood sample was collected. Electrolytes were measured based on ion-selective electrode methods. Fasting blood glucose, and creatinine were measured by spectrophotometric enzyme hexokinase, and Jaffe reaction methods, respectively. The data was entered into Epi-data version 4.6 and analyzed using STATA version 14. Mann–Whitney U-tests and independent t-tests were used for comparison. Multiple logistic regression analysis was done to determine the factors associated with electrolyte imbalances. P-value < 0.05 was considered statistically significant. Result: The overall prevalence of electrolyte imbalance among diabetic patients and controls were 83.07% and 52.31%, respectively. The mean of Na+ and the median level of Mg2+ and Ca2+ were significantly decreased. However, the mean level of Cl− was significantly increased in diabetic patients as compared to control groups. In multivariable logistic regression: alcohol consumption AOR = 3.34 [1.02– 10.9], no formal education AOR = 5.38 [1.14– 25.4], hyperglycemia AOR = 6.32 [2.04– 19.5], and urbanization AOR = 5.6 [1.44– 22.3] showed significant association with electrolyte imbalance. Conclusion: Diabetic patients have more likely to develop electrolyte imbalance than control groups. Diabetic participants showed significantly reduced Na+, Mg2+, and Ca2+ levels and significantly increasing CI− levels when compared to control groups. Hyperglycemia, alcohol drinking habits, urbanization, and no-formal education were statistically significantly associated with electrolyte imbalance. [ABSTRACT FROM AUTHOR]